Compositions for separating molecules

ABSTRACT

The present invention provides compositions for the separation of metals or biomolecules such as polypeptides, nucleic acids, or endotoxins using modified solid supports.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No.60/419,614, filed Oct. 18, 2002.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

Not applicable.

BACKGROUND OF THE INVENTION

The present invention broadly relates to compositions useful inseparating metal ions or other target material including, but notlimited to, polypeptides, nucleic acids, and endotoxins, from non-targetmaterial.

BRIEF SUMMARY OF THE INVENTION

In one aspect, the present invention provides a composition comprising:

wherein R₁ is

-   X is a substituted or unsubstituted alkylene moiety, a substituted    or unsubstituted aralkylene moiety, or a substituted or    unsubstituted arylene moiety;-   R₂ and R₃ are independently selected from R₁, a hydrocarbon moiety,    a substituted hydrocarbon moiety, a halogen atom, a hydrogen atom, a    hydroxy, a thiol, an amine, a silanol bond to the solid support, a    bond to another silane ligand, or O—Si—Y₁Y₂Y₃ wherein Y₁, Y₂ and Y₃    are independently selected from a hydrocarbon moiety or a    substituted hydrocarbon moiety;-   R₄ is selected from a hydrocarbon moiety, a substituted hydrocarbon    moiety, and a hydrogen atom;-   M is a metal ion; and-   n is an integer ≧1.

All publications, patents and patent applications cited herein arehereby incorporated by reference in their entirety. In the case ofconflict between the present disclosure and the incorporatedpublications, the present disclosure should control. It is understoodthat the numerical ranges given herein include all values from the lowervalue to the upper value and that disclosure of the range is a specificdisclosure of all numbers and ranges therein.

These and other aspects of the present invention will be betterappreciated by reference to the following drawings and detaileddescription.

DETAILED DESCRIPTION OF THE INVENTION

The compositions of the present invention comprise modified solidsupports. Conveniently, the compositions of the present invention may beprovided as kits that may be used to separate target material fromnon-target material.

The compositions of the present invention include nitrilotriacetic acid(NTA)-modified solid supports and metal-modified solid supports.Suitable solid supports for making the modified solid supports of thepresent invention include, without limitation, gels or hard supportmaterial, agarose, polyacrylamide, cellulose, plastics, polysaccharides,nylon, polystyrene, latex methacrylate, silica, aluminum oxide,electrodes, membranes and derivatives thereof.

Suitable silica solid supports include, but are not limited to,siliceous oxide, magnetic silica particles, solid silica such as glassor diatomaceous earth and the like, or a combinations of silicamaterials (see, e.g., preparation of silica discussion in Kurt-OthmerEncyclopedia of Chemical Technology, Vol. 21, 4th ed., Mary Howe-Grant,ed., John Wiley & Sons, pub., 1997, p. 1021.) As discussed in theexamples below, suitable silica gels are available commercially fromsuppliers such as Silicycle (Quebec City, Canada), J. T. Baker(Phillipsberg, N.J.), and Sigma-Aldrich, (St. Louis, Mo.). Suitablesilica gels for the compositions and methods of the invention arefurther described in the examples below. Other suitable silica supportsinclude crystalline or vitreous silicas, such as quartz, vitreoussilica, controlled pore glass particles, and glass fibers.

Silica gel may be characterized by pore diameter, particle size, orspecific surface area. Suitable silica gels have a pore diameter fromabout 30 to about 1000 Angstroms, a particle size from about 2 to about300 microns, and a specific surface area from about 50 m²/g to about1000 m²/g. Suitable silica gels include, for example, those having apore diameter of about 40 Angstroms, about 60 Angstroms, and about 150Angstroms; those having a particle size of about 2 to about 25 microns,about 5 to about 25 microns, about 15 microns, about 63 to about 200microns and about 75 to about 200 microns; and those having a specificsurface area of about 300 m²/g, about 500 m²/g, about 550 m²/g, about675 m²/g, and about 750 m²/g.

Conveniently, a solid support according to the present invention maycomprise magnetic silica particles. Magnetic silica particles comprise asuperparamagnetic core coated with a hydrous siliceous oxide adsorptivesurface (i.e. a surface having silanol or Si—OH groups). Suitable,commercially available magnetic silica particles include MagneSil™particles available from Promega Corporation (Madison, Wis.). Thepreparation of magnetic silica particles suitable for use as a supportaccording to the present invention is described in U.S. Pat. No.6,296,937.

Suitable cellulose supports include, but are not limited to,nitrocellulose and cellulose acetate.

Suitable membranes include, but are not limited to, glass fibermembranes impregnated with silica.

Suitable aluminum oxide solid supports include, but are not limited to,Brockmann aluminum oxides that are about 150 mesh and 58 angstroms.

The solid support, as described herein, suitably includes at least onefree hydroxyl group so that when the solid support is contacted with anaminosilane compound, the silicon atom of the aminosilane compound iscovalently bound to the solid support by at least one silanol bond toform an amine-modified solid support.

Aminosilane compounds are commercially available through suppliers suchas United Chemical Technologies, Inc. (Bristol, Pa.). Suitableaminosilane compounds have the general formula

wherein X is an alkylene moiety of up to 20 carbon atoms that may besaturated, unsaturated, branched, linear, or cyclic, for example,methylene, ethylene propylene, nonylene, or an aralkylene moiety of upto 20 carbon atoms in which the alkyl portion may be saturated,unsaturated, branched, linear, or cyclic or an arylene moiety of up to20 carbon atoms, and wherein X may be unsubstituted or substituted asdefined below with respect to hydrocarbon moiety;

-   R₁ is a hydrocarbon moiety, or a substituted hydrocarbon moiety;-   R₂ and R₃ are independently selected from R₁, a hydrocarbon moiety,    a substituted hydrocarbon moiety, a halogen atom, a hydrogen atom, a    hydroxy, a thiol, an amine, a silanol bond to the solid support, a    bond to another silane ligand, or O—Si—Y₁Y₂Y₃, wherein Y₁, Y₂ and Y₃    are independently selected from a hydrocarbon moiety, or a    substituted hydrocarbon moiety; and-   R₄ is a hydrocarbon moiety, a substituted hydrocarbon moiety, or a    hydrogen atom.

The term “hydrocarbon moiety” as used herein refers to an alkyl group ofup to 20 carbon atoms (i.e., alkanes, alkenes or alkynes) that may besaturated, unsaturated, branched, linear, or cyclic; or an aralkyl groupof up to 20 carbon atoms in which the alkyl portion may be saturated,unsaturated, branched, linear or cyclic; or an aryl group of up to 20carbon atoms. Suitably, the hydrocarbon moiety has from 2 to 15 carbonatoms, or from 5 to 10 carbon atoms. A “substituted hydrocarbon moiety”refers to a hydrocarbon moiety, as defined herein, in which at least onecarbon atom is substituted with an oxygen, a sulfur, or a nitrogen atom.The substituent may be, for example, oxo, alkoxy, alkoxycarbonyl,hydroxy, esters, thioethers, amino, alkylamine, or carbamoyl.

Examples of suitable aminosilane compounds useful in the practice of thepresent invention include, but are not limited, to aminopropylsilane,propylethylenediaminesilane,N-[3-(trimethoxysilyl)propyl]ethylenediamine, andN-[3-(trimethoxysilyl)propyl]diethylenetriamine.

An NTA-modified solid support, as described herein, may be produced bycontacting a solid support having a free —NH₂ moiety to form an amidebond between nitrilotriacetic acid and the amine group of the support.Nitrilotriacetic acid acts as a chelating agent capable of formingstable complexes with polyvalent metal ions.

Any solid support is acceptable provided that it has an amine moietythat can be modified or can be made to contain a modifiable amine group.Suitable solid supports for use in the manufacture of NTA-modified solidsupports have a plurality of free NH₂ moieties. One skilled in the artwould be able to attach a free amine functionality to a solid support bychemically modifying the surface of the solid support. See, e.g., GregT. Hermason, A. Krishna Mallia, Paul K. Smith, Immobilized AffinityLegand Techniques, Academic Press (1992). In addition, suitable solidsupports with free NH₂ moieties capable of binding to the NTA to form anNTA-modified solid support according to the present invention arecommercially available. These include, but are not limited to,agarose-based supports sold by Sigma-Aldrich Inc. (St. Louis, Mo.);latex-based supports sold by International Dynamics Corporation,(Longwood, Fla.); polystyrene-based supports sold by Bangs LaboratoriesInc., (Fishers, Ind.); Spherotech, (Libertyville, Ill.); and DynalBiotech, (Lake Success, N.Y.).

In another aspect, the present invention provides metal-modified solidsupports. The metal-modified solid support, as described herein, may beproduced by contacting the NTA-modified solid support described abovewith a metal ion solution to form the metal-modified solid support. Themetal ion solution may be comprised of metal ion salts, wherein thesalts include, but are not limited to chloride, sulfate, phosphate,acetate, carbonate, citrate, acetylacetonate, bromide, fluoride, iodide,nitrate and oxalate salts. The metal concentration may be from less thanabout 10⁻⁶ M to about 1 M. Typically, the metal ion concentration insolution may be in the range of about 0.1 M to about 1 M. It isenvisioned that the metal ion solution may be composed of only one metalion or a mixture of different metals. Suitably, a tetradentate complexmay be formed between the metal ion and the NTA-modified solid support.See, e.g., New multidentate ligands. XV. Chelating tendencies ofdiglycine-N,N-diacetic acid, triglycine-N,N-diacetic acid, andtetraglycine-N,N-diacetic acid, Inorganic Chemistry (1974), 13(3),550–9.

By a “metal ion” it is meant any metal with a oxidation state between +1and +6. Suitably, the metal may be nickel, copper, cobalt, iron, zinc,or gallium. Additionally the following metal ions are consideredsuitable for the present invention: iron (III), copper (II), cobalt(II), nickel (II), zinc (II), cerium (III), magnesium (II), calcium(II), galium (III), chromium (III), indium (III), lanthanum (III),lutetium (III), scandium (III), thallium (III), ytterbium (III), thorium(IV), uranate (II) silver (I), gold (I) and copper (I). One skilled inthe art would be able to select a suitable metal depending on thematerial to be separated. Also, it is envisioned that the bound metalions may be stripped from the metal-modified support with a chelatingagent, such as ethylene diamine tetraacetic acid (EDTA), thereforeallowing the regeneration of the NTA-modified solid supports.

As one of skill in the art will appreciate, the NTA-modified andmetal-modified solid supports may be used in a variety of applicationsas described herein.

The NTA-modified solid support may be used to prepare chelatingimmuno-stimulating complexes using metal chelating approaches asdescribed in U.S. Pat. No. 6,428,807.

It is also envisioned that the NTA-modified solid support may be used toremove toxic metals from drinking water, the environment, or blood ofindividuals with diseases caused by metal exposure. For example, theNTA-modified solid may be used to remove and/or recover potentiallyharmful or toxic metals, such as aluminum, arsenic, bismuth, antimony,excess calcium, excess iron, gold, zinc, magnesium, mercury, cadmium,lead, copper and silver, from industrial waste waters and watersdestined for human consumption.

The NTA-modified solid supports of the present invention may also beuseful in a number of other applications in which it is desirable toextract, deactivate or remove metals from fluids, e.g., removing calciumfrom plasma to convert the plasma to serum, or wiping up spills ofradioactive metallic ions in laboratories. The NTA-modified supports maybe employed to remove toxic metals from individuals with lead or mercurypoisoning.

Interference with or depletion of certain metal ions has been reportedas having a role in health conditions. Accordingly, the NTA-modifiedsolid supports of the present invention may be used as a diagnostic toolfor detecting and extracting metal-associating molecules indicative ofthe disease state or predisposition to a disease.

It is envisioned that metal-modified solid supports may be used toseparate target material (e.g., polypeptides or nucleic acids) fromnon-target material in a starting solution. The metal-modified solidsupport may be used alone, or in conjunction with other purificationmethods, including methods using an amine-modified solid support.

The metal-modified solid support may be utilized in separatinghis-tagged polypeptides from non-target material in a starting solution.The metal-modified solid support may also be used in removal ofendotoxins from a starting solution. Suitably, the term endotoxin refersto the lipopolysaccharide complex associated with the outer membrane ofGram-negative bacteria such as E. coli, Salmonella, Shigella,Pseudomonas, Neisseria, Haemophilus, or any other endotoxin-producingpathogens.

The metal-modified solid support may also be used in the identificationof low-abundance proteins, identification of membrane proteins andphosphorylated proteins. The metal-modified solid support may also beused in the identification and quantitation of polypeptide-polypeptideinteractions where the general classes of suitable detectable moiety orlabel includes but is not limited to a dye, a fluorophore, ananoparticle that may be generally attached anywhere on the targetmaterial or specifically attached to the end of the target material(i.e., N-termini, C-termini or both N-termini and C-termini of peptides)to be detected and quantified.

The metal-modified solid support may also be used in isolatingpolypeptide-polypeptide complexes; screening for polypeptide function;isolating antibodies, antigens and antibody-antigen complexes;quantitating affinity-tagged polypeptides; diagnostic screening fordiseases; antibody screening; antagonist and agonist screening fordrugs; reporter gene assays; producing polypeptide expression libraries,producing polypeptide libraries from cells; producing polypeptidemicroarrays; screening genetically engineered enzymes; or co-isolatinginteracting molecules (i.e., co-factors).

Metal-modified solid supports of the present invention will be useful instudies of polypeptide-polypeptide interactions. The metal-modifiedsolid support may be used to reduce endotoxin concentrations insolution. The metal-modified solid support may be used to separatephosphorylated proteins from a starting solution.

The metal-modified solid support may be utilized in many applications,including, but not limited to, tissue profiling and cell profiling.

The following examples illustrate procedures for practicing theinvention. Those skilled in the art of this invention will appreciatethat the detailed description of the invention is meant to be exemplaryonly and should not be viewed as limiting the scope of the invention.

EXAMPLES Example 1

Preparation Of Metal-Modified3-[[[Bis(carboxymethyl)amino]acetyl]-amino]propyl Silica MagneticParticles.

a) Preparation of 3-Aminopropyl-Modified Magnetic Silica Particles.

3-Aminopropyl-modified magnetic silica particles were prepared asfollows. A 50-ml aliquot of 3-aminopropyltrimethoxysilane was added to astirred solution of methanol (900 mL) followed by addition of water (50mL). The mixture was added to 100 g of magnetic silica particles (MP-50,W. R. Grace, Columbia, Md.). The particles were kept in suspension for 4hr at room temperature using intermittent agitation. The residualmethanol/silane/water solution was removed and the support particleswere washed with 3×1.2 L of water then resuspended in 1 L of methanol.The 3-aminopropyl-modified magnetic silica particles were collected byfiltration and dried under vacuum. Elemental analysis confirmed thecomposition of the 3-aminopropyl-modified magnetic silica particles: C,0.75; H, 0.64; N, 0.30.

b) Preparation of 3-[[[Bis(carboxymethyl)amino]acetyl]amino]-propylMagnetic Silica Particles.

3-[[[Bis(carboxymethyl)amino]acetyl]amino]-propyl magnetic silicaparticles were made by first suspending 3-aminopropyl-modified magneticsilica particles (100 g), prepared as described above, inN,N-dimethylacetamide (600 mL), adding triethylamine (31 ml, 210mmoles), and mixing thoroughly. 200 mmoles of2,6-diketo-N-carboxymethyl-morpholine (prepared according to U.S. Pat.No. 3,621,018) in 400 ml of N,N-dimethylacetamide was added and theresulting mixture was kept in suspension for 4 hr at room temperature.The unreacted N,N-dimethylacetamide/anhydride/triethylamine solution wasremoved and the particles were washed with 3×1.2 L of water. Elementalanalysis confirmed the composition of3-[[[bis(carboxymethyl)amino]acetyl]amino]-propyl-modified magneticsilica particles: C, 1.06; H, 0.61; N, 0.17.

c) Preparation of Nickel (II)3-[[[Bis(carboxymethyl)amino]-acetyl]amino]propyl Magnetic SilicaParticles.

Nickel (II) 3-[[[bis(carboxymethyl)amino]-acetyl]amino]-propyl magneticsilica particles were prepared by suspending 100 grams of3-[[[bis(carboxymethyl)amino]-acetyl]amino]-propyl magnetic silicaparticles, prepared as described above, in a 250 mM nickel (II) chloridesolution (1 L) for 4 hours at room temperature. The excess nickelsolution was removed and the resulting solid support was washed withfive times with water.

Modified particles similar to those described above in Example 1(a)–(c)were prepared using starting particles other than magnetic silicaparticles from W. R. Grace. Other silica gels that have been used insteps (a)–(c) were supplied by: Sigma-Aldrich Corp (St. Louis, Mo.)(23,681-0, 23,682-9, and 23,684-5); Silicyle Inc. (Quebec, Calif.)(S10030M, 10040M, 100300T, S10040T, and R10030M); or J. T. Baker(Philipsburg, N.J.) (7314-02 and 7315-20). The commercial silica gelscontained particles having diameters in the range of about 5 to about500 microns, and pore sizes in the range of about 40 to about 1000Angstroms.

d) Preparation of Colbalt (II)3-[[[Bis(carboxymethyl)amino]-acetyl]amino] propyl Magnetic SilicaParticles.

Cobalt (II) 3-[[[bis(carboxymethyl)amino]-acetyl]amino]-propyl magneticsilica particles were prepared by suspending 100 mg of a3-[[[bis(carboxymethyl)amino]-acetyl]amino]-propyl magnetic silicaparticles, prepared as described above, in a 250 mM cobalt (II) chloridesolution for two minutes at room temperature. The excess cobalt solutionwas removed and the resulting magnetic silica particles were washed 5times with water.

e) Preparation of Copper (II)3-[[[Bis(carboxymethyl)amino]-acetyl]amino] propyl Magnetic SilicaParticles.

Copper (II) 3-[[[bis(carboxymethyl)amino]-acetyl]amino]-propyl magneticsilica particles were prepared by suspending 100 mg of a3-[[[bis(carboxymethyl)amino]-acetyl]amino]-propyl magnetic silicaparticles, prepared as described above, in a CuCl₂ (250 mM) solution fortwo minutes at room temperature. The copper solution was removed and theresulting magnetic silica particles were washed three times with water.

f) Preparation of Zinc (II)3-[[[Bis(carboxymethyl)amino]-acetyl]amino]propyl Magnetic SilicaParticles.

Zinc (II) 3-[[[bis(carboxymethyl)amino]-acetyl]amino]-propyl magneticsilica particles were prepared by suspending 100 mg of a3-[[[bis(carboxymethyl)amino]-acetyl]amino]-propyl magnetic silicaparticles, prepared as described above, in a ZnCl₂ (250 mM) solution fortwo minutes at room temperature. The zinc solution was removed and theresulting magnetic silica particles were washed three times with water.

Example 2

Preparation of Metal-Modified 3-[[[Bis(carboxymethyl)amino]acetyl]-amino]propyl Silica Gel.

(a) Preparation Of 3-Aminopropyl-Modified Silica Gel.

3-Aminopropyltrimethoxysilane (125 mL) was added to a stirred solutionof methanol (2000 mL) followed by addition of water (125 mL). Thismixture was added to 250 g of silica gel (S10040T, 1000 angstom,Silicycle, Inc, Quebec, Canada) and the resulting mixture was kept insuspension for 4 hr at room temperature. After allowing the resin tosettle the residual methanol/silane/water solution was decanted, theparticles were washed with water (3×2.5 L) and resuspended in 2 L ofmethanol. The aminosilane-modified solid support was collected byfiltration and dried under vacuum. Elemental analysis confirmed thecomposition of aminopropyl-modified solid support: C, 0.46; H, 0.30; N,0.19.

(b) Preparation of 3-[[[Bis(carboxymethyl)amino]acetyl]amino]-propylSilica Gel.

3-Aminopropyl-modified solid support (100 g) prepared as described abovewas suspended in N,N-dimethylacetamide (100 mL) and triethylamine (31mL, 210 mmoles) was added to the mixture. This suspension was mixedthoroughly then 200 mmoles of 2,6-diketo-N-carboxymethylmorpholine(prepared according to U.S. Pat. No. 3,621,018, the contents of whichare incorporated herein in its entirety) in 400 mL ofN,N-dimethylacetamide was added and the resulting mixture was kept insuspension for 4 hr at room temperature. The unreactedN,N-dimethylacetamide/anhydride/triethylamine solution was removed andthe solid support was washed with 4×1.2 L of water. Elemental analysisconfirmed the composition of 3-[[[bis(carboxymethyl)amino]acetyl]amino]propyl solid support: C, 0.94; H, 0.32; N, 0.28.

(c) Preparation of Nickel (II)3-[[[Bis(carboxymethyl)amino]acetyl]-amino]propyl Silica Gel.

A portion of 3-[[[bis(carboxymethyl)amino]acetyl]amino]propyl solidsupport prepared as described above was suspended in 250 mM nickel (II)chloride solution for 4 hr at room temperature. The excess nickelsolution was removed and the resulting solid support was washed 5 timeswith water.

Example 3

Preparation of Propylethylenediamine-Modified Magnetic Silica Particles.

N-[3-(Trimethoxysilyl)propyl]ethylenediamine (2 mL) was added to astirred solution of magnetic silica particles (2 g) in 95% methanol (8mL). The resulting mixture was kept in suspension for 4 hr at roomtemperature. The residual methanol/silica solution was removed and theparticles were washed with methanol (5×40 mL) and dried under vacuum.Elemental analysis confirmed the composition ofaminopropylethylenediamine-modified silica magnetic solid support: C,0.97; H, 0.70; N, 0.45.

Example 4

Preparation of Propylethylenediamine-Modified Silica Gel.

N-[3-(Trimethoxysilyl)propyl]ethylenediamine (2 mL) was added to astirred solution of silica particles (1.0 g of Davisil, grade 644 silicagel, 100–200 mesh, 150 A pore size) in 95% methanol (8 mL). Theresulting mixture was kept in suspension for 4 hr at room temperature.The residual methanol/silica solution was removed and the particles werewashed with methanol, 5×40 mL, and dried under vacuum. Elementalanalysis confirmed the composition ofaminopropylethylenediamine-modified silica solid support: C, 5.82; H,1.49; N, 2.44.

The foregoing description of the invention is exemplary for purposes ofillustration and explanation. It will be apparent to those skilled inthe art that changes and modifications are possible without departingfrom the spirit and scope of the invention. It is intended that thefollowing claims be interpreted to embrace all such changes andmodifications.

1. A composition comprising:

wherein R₁ is

X is a substituted or unsubstituted alkylene moiety, a substituted orunsubstituted aralkylene moiety, or a substituted or unsubstitutedarylene moiety; R₂ and R₃ are independently selected from R₁, ahydrocarbon moiety, a substituted hydrocarbon moiety, a halogen atom, ahydrogen atom, a hydroxy, an alkoxy, a thiol, an amine, a silanol bondto the solid support, a bond to another silane ligand, or O—Si—Y₁Y₂Y₃,wherein Y₁, Y₂ and Y₃ are independently selected from a hydrocarbonmoiety or a substituted hydrocarbon moiety; R₄ is a hydrocarbon moiety,a substituted hydrocarbon moiety, or a hydrogen atom; M is a metal ion;and n is an integer ≧1.
 2. The composition of claim 1, wherein the metalion has an oxidation state of +1 to +6.
 3. The composition of claim 2,wherein the metal ion is nickel, copper, cobalt, iron, zinc or gallium.4. The composition of claim 1, wherein X is a saturated alkylene groupor a substituted saturated alkylene group.
 5. The composition of claim1, wherein X is a saturated alkylene group of up to 10 carbon atoms or asubstituted saturated alkylene group of up to 10 carbon atoms.
 6. Thecomposition of claim 5, wherein X is —(CH₂)₃—.
 7. The composition ofclaim 1, wherein the solid support is selected from the group consistingof silica gels, siliceous oxides, solid silicas, magnetic silicaparticles, crystalline silicas, vitreous silicas, aluminum oxide, andcombinations thereof.
 8. A composition comprising

wherein R₂ and R₃ are independently selected from R₁, a hydrocarbonmoiety, a substituted hydrocarbon moiety, a halogen atom, a hydrogenatom, a hydroxy, an alkoxy, a thiol, an amine, a silanol bond to thesolid support, a bond to another silane ligand, or O—Si—Y₁Y₂Y₃, whereinY₁, Y₂ and Y₃ are independently selected from a hydrocarbon moiety or asubstituted hydrocarbon moiety.
 9. A method of making a modified solidsupport comprising:

wherein R₁ is

X is a substituted or unsubstituted alkylene moiety, a substituted orunsubstituted aralkylene moiety, or a substituted or unsubstitutedarylene moiety; R₂ and R₃ are independently selected from R₁, ahydrocarbon moiety, a substituted hydrocarbon moiety, a halogen atom, ahydrogen atom, a hydroxy, a thiol, an amine, a silanol bond to the solidsupport, a bond to another silane ligand, or O—Si—Y₁Y₂Y₃, wherein Y₁, Y₂and Y₃ are independently selected from a hydrocarbon moiety or asubstituted hydrocarbon moiety; R₄ is a hydrocarbon moiety, asubstituted hydrocarbon moiety, or a hydrogen atom; and n comprises aninteger ≧1, comprising: (a) contacting a solid support with anaminosilane compound to form a first complex, having a silanol bondbetween the solid support and the aminosilane compound; (b) contactingthe first complex with nitrilotriacetic acid to form a modified solidsupport having an amide bond between the nitrilotriacetic acid and thefirst complex; and (c) contacting the modified solid support with ametal ion solution to form a metal-modified solid support.
 10. Themethod of claim 9, wherein the aminosilane compound contains a saturatedalkylene chain or a substituted saturated alkylene chain.
 11. The methodof claim 9, wherein the aminosilane compound is aminopropylsilane. 12.The method of claim 9, wherein the aminosilane compound is selected fromthe group consisting of aminopropylsilane, propylethylenediaminosilane,and aminopropyltrimethoxysilane compounds.
 13. The method of claim 9,wherein the solid support is selected from the group consisting ofsilica gels, siliceous oxides, solid silicas, magnetic silica particles,crystalline silicas, vitreous silicas, aluminum oxide, and combinationsthereof.
 14. The method of claim 9, wherein the metal ion has anoxidation state of +1 to +6.
 15. The method of claim 14, wherein themetal ion is nickel, copper, cobalt, iron, zinc or gallium.
 16. Acomposition prepared by the process of claim 9.